| Hepatocytes are the major functional cellular constituent comprising liver tissue. A critical problem is the lack of available culture systems supporting extended survival and continued function of hepatocytes outside of their native liver tissue. Several hepatocyte culture systems support short-term hepatocyte culture but these cells quickly de-differentiate and lose their metabolic functions. Other culture systems do not provide appropriate substrates for adhesion. There is also a lack of appropriate materials and patterning designs that will support hepatocyte adhesion, proliferation, and function.;The overall goal of the study was to test the hypothesis that a hepatocyte culture model with the appropriate material and surface design which simulates the in vivo liver microenvironment will support hepatocyte adhesion and proliferation for up to one week in culture. The three-dimensional organizational structure of hepatocytes in liver tissue could be mimicked by utilizing micropatterned agarose with a coating of crosslinked collagen. Hepatocyte adhesion, morphology, behavior, and function were dependent on the specific chemical characteristics of the culture substrate and on surface topography. Controlling the cell microenvironment by micropatterning provided a means to modulate cell differentiation, cell-cell communication, and cell signaling pathways.;Establishment of a hepatocyte model may be useful to better understand mechanisms influencing liver damage and disease by defining cell signaling pathways regulating genes responsible for cellular and metabolic functions. Hepatocytes continued to respond to externally applied stimuli by inducing production of inflammatory mediators and oxidative stress. Therefore, this model facilitates the study of hepatocytes in response to a range of agents with anti-inflammatory and anti-oxidant properties. This model may also be useful for screening and identification of hepatoprotective and hepatotoxic agents, drug discovery, assessment of pre-clinical safety and efficacy, as well as development of therapeutic agents to protect the liver.;This study has successfully utilized an innovative combination of a biomaterial and surface patterning for hepatocyte culture. Culture of hepatocytes in a controlled environment simulating that found in liver maintains the integrity of hepatocytes and takes into account the native liver milieu and cell configuration. |
